OBJECTIVE To establish a combined pharmacokinetic(PK)-pharmacodynamic(PD)model for knee osteoarthritis(KOA)of crossbow drug microemulsion multi-components(benzoylmesaconine,benzoylhypacoitine,mesaconitine,periplocin,neo-chlorogenic acid,vanillic acid,chlorogenic acid),and elucidate the dynamic changes in the KOA rats and the interrelation with the e-lapsed efficacy of the drug.METHODS A KOA rat model was induced by 4%papain;the PK process of crossbow medicine microe-mulsion components in rat synovial fluid was analyzed by UPLC to establish a PK model;the contents of MMP-3,MMP-13,TNF-α and IL-1β in KOA rats at different time points after administration were determined by ELISA analysis to establish a PD model;Phoe-nix WinNonlin software was used to fit the PK and PD data to obtain a PK-PD model.RESULTS PK results showed that the multi-components of the microemulsion were slowly absorbed in the joint cavity and gradually reached the peak value within 3-5 h.The Cmax of benzoylmesaconine,benzoylhypacoitine mesaconitine,periplocoside,neochlorogenic acid,vanillic acid and chlorogenic acid were 1.23,1.48,1.62,4.67,0.93,1.25 and 2.35 μg·mL-1,respectively;the area under the drug-time curve(AUC0-11)was 2.58,4.04,3.54,12.15,2.51,2.41 and 4.11 h·μg·mL-1,respectively.PD results showed that at different time points after adminis-tration,the contents of MMP-3,IL-1β,TNF-α,and MMP-13 decreased to varying degrees,among which MMP-3 decreased insig-nificantly,with significant differences only at 6 h;the contents of the remaining IL-1β,TNF-α,and MMP-13 decreased significantly(P<0.05,P<0.01),and showed the phenomenon of lagged efficacy;the PK-PD binding model showed that the drug concentration of the multi-component drug in the crossbow medicine microemulsion could be well fitted with its drug efficacy data.CONCLUSION The established PK-PD binding model can predict the drug efficacy changes after administration,and provides a corresponding refer-ence for the crossbow medicine microemulsion treatment of KOA.

OBJECTIVE To establish a combined pharmacokinetic(PK)-pharmacodynamic(PD)model for knee osteoarthritis(KOA)of crossbow drug microemulsion multi-components(benzoylmesaconine,benzoylhypacoitine,mesaconitine,periplocin,neo-chlorogenic acid,vanillic acid,chlorogenic acid),and elucidate the dynamic changes in the KOA rats and the interrelation with the e-lapsed efficacy of the drug.METHODS A KOA rat model was induced by 4%papain;the PK process of crossbow medicine microe-mulsion components in rat synovial fluid was analyzed by UPLC to establish a PK model;the contents of MMP-3,MMP-13,TNF-α and IL-1β in KOA rats at different time points after administration were determined by ELISA analysis to establish a PD model;Phoe-nix WinNonlin software was used to fit the PK and PD data to obtain a PK-PD model.RESULTS PK results showed that the multi-components of the microemulsion were slowly absorbed in the joint cavity and gradually reached the peak value within 3-5 h.The Cmax of benzoylmesaconine,benzoylhypacoitine mesaconitine,periplocoside,neochlorogenic acid,vanillic acid and chlorogenic acid were 1.23,1.48,1.62,4.67,0.93,1.25 and 2.35 μg·mL-1,respectively;the area under the drug-time curve(AUC0-11)was 2.58,4.04,3.54,12.15,2.51,2.41 and 4.11 h·μg·mL-1,respectively.PD results showed that at different time points after adminis-tration,the contents of MMP-3,IL-1β,TNF-α,and MMP-13 decreased to varying degrees,among which MMP-3 decreased insig-nificantly,with significant differences only at 6 h;the contents of the remaining IL-1β,TNF-α,and MMP-13 decreased significantly(P<0.05,P<0.01),and showed the phenomenon of lagged efficacy;the PK-PD binding model showed that the drug concentration of the multi-component drug in the crossbow medicine microemulsion could be well fitted with its drug efficacy data.CONCLUSION The established PK-PD binding model can predict the drug efficacy changes after administration,and provides a corresponding refer-ence for the crossbow medicine microemulsion treatment of KOA.

Objective:To explore the protective effect of astaxanthin on acute liver injury induced by α-amanitin in mice.Methods:In June 2023, 42 healthy SPF male Kunming mice were selected. The mice were divided into blank control group, model (0.45 mg/kg α-amanitin) group, olive oil (10 ml/kg olive oil) group, low dose (20 mg/kg) astaxanthin group, medium dose (40 mg/kg) astaxanthin group, high dose (80 mg/kg) astaxanthin group and silybin (20 mg/kg) group by random number table method. Each group had 6 animals. Mice in the blank control group were intraperitoneally injected with 10 ml/kg normal saline, and mice in the other group were injected with α-amanitin. After that, the blank control group and model group were infused with 10 ml/kg normal saline, olive oil group and astaxanthin groups were given olive oil and astaxanthin according to dose by gavage, and silybin group was injected with silybin by dose. The drug was administered once every 12 h for a total of 4 doses. After 60 h, the mice were killed, the liver weight was weighed, and the liver index was calculated. The contents of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in serum of mice were detected, and the contents of superoxide dismutase (SOD), reduced glutathione (GSH), catalase (CAT), malondialdehyde (MDA) in liver tissues were also detected. One-way analysis of variance (ANOVA) was used to compare the difference of indexes among each group, and pairwise comparison was performed by Dunnett- t test. Results:The mice in the blank control group had smooth hair color, good spirit and normal behavior, while the mice in the other groups showed varying degrees of retardation and decreased diet, and no death occurred in each group. Body mass[ (26.67±1.51) g] and liver mass[ (1.23±0.14) g] in model group were significantly lower than those in blank control group [ (33.50±2.43) g and (1.87±0.16) g], and the differences were statistically significant ( P<0.05). The liver index [ (5.39±0.32) %, (5.83±0.30) %, (5.75±0.24) % and (5.78±0.16) %] in low, medium and high dose astaxanthin groups and silybin group were significantly higher than those in model group [ (4.61±0.12) %], and the differences were statistically significant ( P<0.05). Serum ALT and AST contents in model group [ (153.04±13.96) U/L and (59.08±4.03) U/L] were significantly higher than those in blank control group [ (13.77±1.29) U/L and (10.21±0.35) U/L], and the differences were statistically significant ( P<0.05). The contents of CAT, GSH and SOD in liver tissues of model group [ (9.40±2.23) U/mgprot, (3.09±0.26) μmol/gprot and (48.94±3.13) U/mgprot] were significantly lower than those of blank control group [ (26.36±2.92) U/mgprot, (6.76±0.71) μmol/gprot and (89.89±4.17) U/mgprot], the differences were statistically significant ( P<0.05). MDA content[ (6.33±0.24) nmol/mgprot] in liver tissue of model group was significantly higher than that of blank control group [ (0.91±0.21) nmol/mgprot], and the difference was statistically significant ( P<0.05). The CAT contents[ (18.64±1.76) U/mgprot, (18.20±1.76) U/mgprot, and (15.54±1.36) U/mgprot] in liver tissues of low, medium and high dose astaxanthin groups were significantly higher than those of model group, with statistical significances ( P<0.05). Compared with model group, SOD contents[ (72.16±7.44) U/mgprot, (93.18±5.28) U/mgprot, (103.78±7.07) U/mgprot, and (96.60±7.02) U/mgprot] in liver tissues of mice in low, medium and high dose astaxanthin groups and silybin group were significantly increased ( P<0.05), MDA contents [ (4.30±0.84) U/mgprot, (3.66±0.28) U/mgprot, (2.96±0.29) U/mgprot, and (2.88±0.39) U/mgprot] were significantly decreased ( P<0.05). Compared with model group, GSH content [ (7.90±1.25) μmol/gprot] in high dose astaxanthin group was significantly increased ( P<0.05) . Conclusion:Astaxanthin may alleviate acute liver injury induced by α-amanitin by alleviating oxidative stress in mice liver.

Objective:To explore the protective effect of astaxanthin on acute liver injury induced by α-amanitin in mice.Methods:In June 2023, 42 healthy SPF male Kunming mice were selected. The mice were divided into blank control group, model (0.45 mg/kg α-amanitin) group, olive oil (10 ml/kg olive oil) group, low dose (20 mg/kg) astaxanthin group, medium dose (40 mg/kg) astaxanthin group, high dose (80 mg/kg) astaxanthin group and silybin (20 mg/kg) group by random number table method. Each group had 6 animals. Mice in the blank control group were intraperitoneally injected with 10 ml/kg normal saline, and mice in the other group were injected with α-amanitin. After that, the blank control group and model group were infused with 10 ml/kg normal saline, olive oil group and astaxanthin groups were given olive oil and astaxanthin according to dose by gavage, and silybin group was injected with silybin by dose. The drug was administered once every 12 h for a total of 4 doses. After 60 h, the mice were killed, the liver weight was weighed, and the liver index was calculated. The contents of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in serum of mice were detected, and the contents of superoxide dismutase (SOD), reduced glutathione (GSH), catalase (CAT), malondialdehyde (MDA) in liver tissues were also detected. One-way analysis of variance (ANOVA) was used to compare the difference of indexes among each group, and pairwise comparison was performed by Dunnett- t test. Results:The mice in the blank control group had smooth hair color, good spirit and normal behavior, while the mice in the other groups showed varying degrees of retardation and decreased diet, and no death occurred in each group. Body mass[ (26.67±1.51) g] and liver mass[ (1.23±0.14) g] in model group were significantly lower than those in blank control group [ (33.50±2.43) g and (1.87±0.16) g], and the differences were statistically significant ( P<0.05). The liver index [ (5.39±0.32) %, (5.83±0.30) %, (5.75±0.24) % and (5.78±0.16) %] in low, medium and high dose astaxanthin groups and silybin group were significantly higher than those in model group [ (4.61±0.12) %], and the differences were statistically significant ( P<0.05). Serum ALT and AST contents in model group [ (153.04±13.96) U/L and (59.08±4.03) U/L] were significantly higher than those in blank control group [ (13.77±1.29) U/L and (10.21±0.35) U/L], and the differences were statistically significant ( P<0.05). The contents of CAT, GSH and SOD in liver tissues of model group [ (9.40±2.23) U/mgprot, (3.09±0.26) μmol/gprot and (48.94±3.13) U/mgprot] were significantly lower than those of blank control group [ (26.36±2.92) U/mgprot, (6.76±0.71) μmol/gprot and (89.89±4.17) U/mgprot], the differences were statistically significant ( P<0.05). MDA content[ (6.33±0.24) nmol/mgprot] in liver tissue of model group was significantly higher than that of blank control group [ (0.91±0.21) nmol/mgprot], and the difference was statistically significant ( P<0.05). The CAT contents[ (18.64±1.76) U/mgprot, (18.20±1.76) U/mgprot, and (15.54±1.36) U/mgprot] in liver tissues of low, medium and high dose astaxanthin groups were significantly higher than those of model group, with statistical significances ( P<0.05). Compared with model group, SOD contents[ (72.16±7.44) U/mgprot, (93.18±5.28) U/mgprot, (103.78±7.07) U/mgprot, and (96.60±7.02) U/mgprot] in liver tissues of mice in low, medium and high dose astaxanthin groups and silybin group were significantly increased ( P<0.05), MDA contents [ (4.30±0.84) U/mgprot, (3.66±0.28) U/mgprot, (2.96±0.29) U/mgprot, and (2.88±0.39) U/mgprot] were significantly decreased ( P<0.05). Compared with model group, GSH content [ (7.90±1.25) μmol/gprot] in high dose astaxanthin group was significantly increased ( P<0.05) . Conclusion:Astaxanthin may alleviate acute liver injury induced by α-amanitin by alleviating oxidative stress in mice liver.

Objective:To investigate the clinical effect of homemade adjustable mirabilite vest in patients with severe acute pancreatitis and supply reference for clinical nursing.Methods:This was a randomized controlled study. One hundred patients with acute severe pancreatitis admitted to Putuo Hospital, Shanghai University of Traditional Chinese Medicine from January 2021 to June 2022 were selected, and were divided into the pocket group and the vest group according to the order of admission with 50 cases in each group. The pocket group used traditional mirabilite bag for external application, the vest group used adjustable mirabilite vest for external application. The other treatment measures were the same for both two group. The comfort degree, itching severity and average length of hospital stay of these two groups were compared.Results:The basic data of the two groups were homogeneous. The difference were not statistically significant( P>0.05). After intervention, the comfort degree of the pocket group was (65.90 ± 7.95) points while the comfort degree of the vest group was (77.04 ± 5.96) points. The difference was statistically significant ( t = 7.93, P<0.01). The degree of pruritus was (12.72 ± 3.95) points in the pocket group and (8.00 ± 1.20) points in the vest group.The difference was statistically significant ( t = 8.08, P<0.05). The mean length of hospital stay in the pocket group was (15.86 ± 5.83) days and (11.02 ± 3.38) days in the vest group. The difference was statistically significant ( t = 5.08, P<0.01). Conclusions:When using topical mirabilite for patients with acute severe pancreatitis, the use of adjustable mirabilite vest can significantly improve patients′ comfort, reduce itching, and reduce the number of hospital days, which has the value of promotion and use.

OBJECTIVE To prepare celastrol -loaded albumin nanoparticles （CLT-AN），and to investigate their activity against rheumatoid arthritis （RA）in vivo . METHODS CLT-AN was prepared by ultrasonic method . The formulation technology was optimized by single -factor test by taking particle size ，polydispersity index （PDI）and stability as indexes ，with the dosage of CLT ， the dosage of soybean oil and the ultrasonic power as factors . The physical and chemical properties of CLT -AN were investigated by transmission electron microscopy （TEM）and laser particle size analyzer ；in vitro stability and release profile were studied . A rat model of adjuvant -induced arthritis was constructed to investigate the effects of CLT -AN on joint swelling ，the levels of serum inflammatory factors [tumor necrosis factor α（TNF-α）and interleukin -1β（IL-1β）] and pathological state of joint tissue . RESULTS The optimized formulation was CLT 6.5 g，soybean oil 45 mg，ultrasonic power 490 W，ultrasonic time 8 min. CLT- AN prepared by the best formulation showed uniform and spherical morphology . Its particle size ，PDI，Zeta potential were （96.8± 1.1）nm，0.174±0.020，and（－18.6±1.7）mV，respectively. The encapsulation efficiency and drug -loading efficiency were （94.61±0.46）% and（2.42±0.21）%. There were no significant changes in particle size ，PDI，Zeta potential and encapsulation efficiency of CLT -AN within 5 days of storage at room temperature . CLT-AN was slowly released in vitro ，and the cumulative release reached 73.56% in 72 h. Compared with CLT ，CLT-AN could significantly inhibit the joint swelling of model rats ，reduced the levels of inflammatory factors TNF -α and IL -1β in serum ，and improved the pathological state of inflammatory joint tissue . CONCLUSIONS CLT-AN prepared by ultrasonic method has the appropriate particle size ，good stability ，significant sustained - release characteristics ，and excellent therapeutic efficacy against RA .

Objective:To investigate the mid-term clinical outcomes of selective column arthrodesis based on the three-column theory in the treatment of malunion of Lisfranc injury.Methods:The 28 patients with malunion of Lisfranc injury were analyzed retrospectively who had been treated by selective column arthrodesis at Department of Orthopedic Surgery, The Second Affiliated Hospital, Zhejiang University School of Medicine from January 2011 to January 2020.They were 18 males and 10 females, with an average age of 37.2 years(from 18 to 65 years). Twelve left and 16 right sides were affected. According to Myerson's three-column classification, one case was medial column injury (type A), 4 ones middle column injury (type B), 7 ones medial plus middle columns injury and 16 ones three-column injury. Medial column arthrodesis was conducted in 7, middle column arthrodesis in 4 and medial plus middle columns arthrodesis in 17. The American Orthopaedic Foot and Ankle Society (AOFAS) midfoot score and visual analogue scale (VAS) were compared between preoperation and the last follow-up to evaluate the improvements in foot function and pain. The operation-related complications were recorded.Results:All patients were followed up for an average of 35.6 months (from 18 to 60 months). The AOFAS midfoot score increased from 43.1±4.1 at pre-operation to 84.1± 7.4 at the last follow-up and the VAS score decreased from 5.7±1.3 at pre-operation to 2.0±0.9 at the last follow-up (both P<0.001). The wounds healed in 28 patients, 3 of whom had postoperative wound exudation but responded to dressing change. There were no such complications as injury to the deep peroneal nerve or deep venous thrombosis. The internal fixation was removed in 5 patients at about one year after arthrodesis. Conclusion:Selective column arthrodesis based on the three-column theory can result in satisfactory med-term clinical outcomes in the treatment of malunion of Lisfranc injury.

Objective:To evaluate the immunological efficacy of a novel DNA vaccine against West Nile virus (WNV) in a mouse model.Methods:A DNA vaccine VRC-prME expressing the precursor membrane (prM) and envelope protein (E) of WNV Xinjiang strain (XJ11129-3) was constructed and its ability to express virus-like particles was verified in vitro. C57BL/6 mice were immunized twice with VRC-prME via intramuscular injection combined with electroporation with an interval of four weeks. Enzyme-linked immunoassay (ELISA) was used to detect serum antibodies after immunization. WNV (NY99 strain) single-round infectious particles were used to detect neutralizing antibodies. Cellular immune responses were analyzed by enzyme-linked immunoblot assay (ELISPOT) and intracellular cytokine staining (ICS). Results:VRC-prME induced a strong Th1-biased antibody response in mice that could cross-neutralize the WNV (NY99 strain) single-round infectious particles two weeks after the boost immunization. Moreover, the vaccine also elicited antigen-specific multifunctional CD8 + T cell responses (IFN-γ, IL-2, TNF-α). Conclusions:The novel DNA vaccine prepared in this study, expressing the prME protein of WNV XJ11129-3 strain, could induce stronger humoral and cellular immune responses in mice, which was worthy of further research and development for the prevention of WNV infection in China.

Objective To study the degradation behavior and mechanical properties of magnesium alloy plate on treatment of tibial fracture in New Zealand rabbits. Methods Thirty-six adult New Zealand rabbits were randomly divided into experimental group (magnesium alloy bone plate group, n=18) and control group (titanium alloy bone plate group, n=18). Tibial fractures in experimental group and control group were fixed with magnesium alloy bone plate and titanium alloy bone plate, respectively. After operation, X-ray, scanning electron microscopy, energy spectrum analysis, weight loss test and four-point bending test were performed in each group to analyze the degradation behavior and mechanical properties of magnesium alloy plate after tibial fracture treatment. Results Magnesium alloy bone plate could be degraded gradually in vivo. The degradation of magnesium alloy bone plate was deepened gradually with the implantation time, and the surface was corroded uniformly. The mechanical properties of magnesium alloy bone plate decreased gradually with the degradation in vivo. Conclusions Magnesium alloy bone plate can degrade gradually with fracture healing in vivo, and its mechanical properties gradually decline, but it can still meet the requirements of fracture internal fixation, and is a kind of good new degradable orthopedic implant material.

OBJECTIVE：To c ompare the difference of volatile oil and fatty oil constituents from Cinnamomum migao in different sources. METHODS ：The steam distillation method and Soxhlet extraction mothod were used to extract volatile oil and fatty oil from C. migao in different sources respectively ，and the extraction rates were calculated ；GC-MS was used to analyze volatile oils and fatty oils constituents from C. migao in different sources. The compounds were searched and matched through NIST 17，WILEY 275 databases and mass spectrometry computer date system. The relative percentage content of each constituent was calculated by peak area normalization method. RESULTS ：The extraction rates of the volatile oils from 4 batches of C. migao in different sources were 3.1％，4.5％，6.2％ and 5.5％，respectively；the extraction rates of the fatty oils from C. migao were 6.2％，8.3％，10.5％ and 9.4％，respectively. A total of 87 constituents were identified in 4 batches of volatile oils of C. migao in different sources ，of which 104 constituents were separated from S 1，67 were identified ，and the relative percentage content was 90.172％；102 constituents were separated from S2，73 were identified ，and the relative percentage contentwas 88.836％；77 constituents were separated from S 3，57 were identified ， with a relative percentage content of 93.972％；87 constituents were separated from S 4，60 were m identified，with a relative percentage content of 95.247％ . Among above 87 constituents，48 were monotyloids and their derivat ives，33 were sesquiterpenoids and their derivatives ，4 were aliphatic and 2 were ketones. There were 44 common constituents from the volatile oil of C. migao in different sources ，all of which were terpenoids. The relative percentage content of S 1-S4 were 38.556％，66.776％，88.886％ and 90.115％，respectively. Among 44 common constituents ，the relative percentage content of which were all greater than 1％ were 1，8-cineole（S2： 6.518％；S4：3.850％；S3：1.655％；S1：1.475％；），4-terpineol（S2：1.591％；S4：1.384％；S3：1.193％；S1：1.182％）， α-terpinenol（S3：8.662％；S4：7.173％；S2：6.503％；S1：4.839 ％），δ-cadinene（S3：8.597％；S4：5.329％；S2：2.677％； S1：2.547％），elemol（S3：4.781％；S2：4.113％；S1：2.568％；S4：1.897％）and γ-eudesmol（S2：4.061％；S3：2.167％；S1： 1.575％；S4：1.197％）. A total of 37 constituents were identified in the 4 batches of fatty oil of the C. migao in different sources ， of which 87 constituents were separated from S 1，34 were identified ，and the relative percentage content was 91.072％；69 constituents were separated from S 2，28 were identified ，and the relative percentage content was 90.527％；63 constituents were separated from S 3，23 were identified ，the relative percentage content was 85.297％；71 constituents were separated from S 4，24 were identified ，with relative percentage content of 91.527％. Among above 37 constituents，there were 21 monoterpenes and their derivatives，2 sesquiterpenes，13 aliphatics，and 1 alkane. There were 20 common constituents in fatty oil from C. migao of different sources ，and the relative percentage content in S 1-S4 were 89.667％，89.595％，84.651％ and 90.972％，respectively. Among 20 common constituents ，the constituents with relative percentage content greater than 1％ were methyl caprate （S4： 59.498％；S1：58.733％；S2：57.552％；S3：26.423％）and methyl dodecanoate （S3：31.434％；S2：26.990％；S1：25.095％； S4：24.334％）. CONCLUSIONS ：There are differences in volatile oil and fatty oil constituents of C. migao from different sources ， and the contents of the same constituent were also different.